Fig. 3: PdCDIE1 interacts with citrus CsHsp70.
From: A conserved fungal effector disturbs Ca2+ sensing and ROS homeostasis to induce plant cell death
a The interaction between PdCDIE1 and CsHsp70 was confirmed by yeast two-hybrid (Y2H) assays. The empty pGADT7 vector and pGADT7-GFP vector served as negative controls, and transformed yeast cells were grown on selective media. EV, empty vector; SD-2, SD/–Leu/–Trp; SD-4 + X-α-gal, SD/–Ade/–His/–Leu/–Trp containing X-α-gal. b PdCDIE1 colocalized with CsHsp70 in plant cell. PdCDIE1-GFP and mCherry-CsHsp70 were co-expressed in N. benthamiana leaves. Bars, 20 μm. c Visualization of protein interactions in planta by bimolecular fluorescence complementation (BiFC) assays. Micrographs show N. benthamiana leaves transiently expressing constructs encoding the indicated fusion proteins. AtUGPase-mCherry was used as a cytoplasm marker. Bars, 20 μm. d, e The interaction between PdCDIE1 and CsHsp70 was confirmed by co-immunoprecipitation (Co-IP) assays. Co-IP assays were performed based on Agrobacterium-mediated transient expression of PdCDIE1 and CsHsp70 in citrus fruit. CBB, Coomassie Brilliant Blue. All these experiments were repeated three times with similar results.
